Aerobic inactivation of Rhizobium meliloti NifA in Escherichia coli is mediated by lon and two newly identified genes, snoB and snoC

Abstract

The Rhizobium meliloti NifA protein is an oxygen-sensitive transcriptional regulator of nitrogen fixation genes. Regulation of NifA activity by oxygen occurs at the transcriptional level through fixLJ and at the posttranslational level through the sensitivity of NifA to oxygen. We have previously reported that the NifA protein is sensitive to oxygen in Escherichia coli as well as in R. meliloti. To investigate whether the posttranslational regulation of NifA is dependent on host factors conserved between R. meliloti and E. coli, we carried out a Tn5 mutagenesis of E. coli and isolated mutants with increased NifA activity under aerobic conditions. Fifteen insertion mutations occurred at three unlinked loci. One locus is the previously characterized lon gene; the other two loci, which we have named snoB and snoC, define previously uncharacterized E. coli genes. The products of snoC and lon affect the rate of NifA degradation, whereas the product of snoB may affect both NifA degradation and inactivation. A snoB lon double mutant showed a higher level of NifA accumulation than did a lon mutant, suggesting that the snoB product affects the ability of NifA to be degraded by a lon-independent pathway. The effects of a snoC mutation and lon mutation were not additive, suggesting that the snoC and lon products function in the same degradative pathway.